| Autor: |
Milenković, Milan, Đajić, Nevena, Krmar, Jovana, Rašević, Marija, Malenović, Anđelija, Otašević, Biljana, Protić, Ana |
| Jazyk: |
angličtina |
| Rok vydání: |
2021 |
| Zdroj: |
13th International Symposium on Pharmaceutical Sciences (ISOPS), June 22-25, 2021, Ankara, Turkey |
| Popis: |
Introduction: Gradient elution HPLC finds its purpose in simultaneous analyses of solutes covering wide range of polarities. However, the instrument related factors, especially dwell volume, are frequently responsible for fizzy transfer and short life cycle of the gradient elution method. Therefore, it is advisable to incorporate dwell volume into the optimization stage and avoid transfer related failures. The chemometric approach would enable selection of optimal chromatographic conditions for different HPLC instruments. The aim of this study was to propose and test this approach in gradient elution method’s development. Materials and Methods: The experiments were carried out on three chromatographic systems (UPLC, UHPLC and HPLC), while the separation was achieved on Kinetex C18 Core-shell column (100 mm × 2.1 mm, 2.6 μm particle size). Design of experiments was constructed in Design-Expert 11.0. Indirect modeling, grid point search and graphical presentations were done in Matlab 7.10.0. Results: Dabigatrane etexilate mesylate and nine structurally related compounds were selected as suitable model mixture due to its complexity and polarity. Method development was supported with experimental design methodology, Placket – Burman for screening and D-optimal design for optimization purposes. Dwell volumes were included in the optimization phase and in this way the same optimal chromatographic conditions for all three instruments were selected. They included 10 mM ammonium acetate buffer with pH set to 4.9 using acetic acid, and acetonitrile. The components of the mobile phase were pumped into chromatographic system with flow rate of 400 μL min-1 in a linear gradient mode: at 0 minutes 24% (v/v) acetonitrile and 76% (v/v) of buffer solution, at 15 minutes 54% (v/v) acetonitrile and 46% (v/v) buffer solution. At 16 minutes the acetonitrile content was back to 24% (v/v) and 76% (v/v) of buffer solution. The re-equilibration time was set to 5 minutes. The examined chromatographic region is graphically presented and optimal conditions are noticed as the cross sections (yellow dots). The method was validated and confirmed its utility on all instruments. Conclusions: The proposed methodology demonstrated its ability to predict joint optimal chromatographic conditions for instruments with different values of dwell volume. The potential was confirmed on complex model mixture and instruments significantly differing in dwell volume values. In this way the gap between developing and routine needs could be overwhelmed, followed by facilitated transfer of methods. ISOPS 13, Abstract Book |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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